CARF regulates the alternative splicing and piwi/piRNA complexes during mouse spermatogenesis through PABPC1

Area of Science:

• Molecular Biology
• Reproductive Biology
• Genetics

Background:

• ADP-ribosylation factor collaborator (CARF) is an RNA-binding protein involved in cell cycle regulation and stem cell maintenance.
• CARF's role in mammalian spermatogenesis remains largely unexplored.

Purpose of the Study:

• To investigate the function of CARF in male reproductive processes.
• To determine the molecular mechanisms underlying CARF's role in spermatogenesis and male fertility.

Main Methods:

• Generated Carf-deficient male mice.
• Utilized immunoprecipitation-mass spectrometry (IP-MS) and RNA sequencing (RNA-seq).
• Analyzed splicing patterns, mRNA targets, and PIWIL1 expression.

Main Results:

• Carf-deficient mice exhibited impaired spermatogenesis and reduced fertility.
• CARF interacts with splicing factors and targets specific mRNAs in spermatocytes.
• CARF deficiency led to aberrant mRNA splicing and altered PIWIL1 expression and pachytene-piRNA ratios.

Conclusions:

• CARF is crucial for regulating alternative splicing during mammalian spermatogenesis.
• CARF plays a vital role in maintaining male fertility, and its disruption leads to infertility.